Protein kinase required for Ras signal transduction
The kinase suppressor of Ras (Ksr), a novel protein kinase involved in the regulation of cell growth and differentiation, provides an important target for therapeutic intervention. The subject compositions also include nucleic acids which encode a Ksr kinase, and hybridization probes and primers capable of hybridizing with a Ksr gene. Such probes are used to identify mutant Ksr alleles associated with disease. The invention includes methods, including phosphorylation and binding assays, for screening chemical libraries for lead compounds for a pharmacological agent useful in the diagnosis or treatment of disease associated Ksr activity or Ksr-dependent signal transduction.
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1. An isolated kinase suppressor of ras (Ksr) protein.
2. An isolated kinase suppressor of ras (Ksr) protein according to claim 1, wherein said protein is mammalian.
3. An isolated kinase suppressor of ras (Ksr) protein according to claim 1, wherein said protein is human.
4. A recombinant isolated Ksr protein produced by a cell comprising a nucleic acid encoding a kinase suppressor of ras (Ksr) according to claim 1 operably linked to a transcription regulatory region not naturally lined to a Ksr-encoding gene.
5. A recombinant isolated Ksr protein according to claim 4, wherein said cell is a mammalian cell, an avian cell, an insect cell, a fungal cell, an amphibian cell or a fish cell.
6. An isolated kinase suppressor of ras (Ksr) protein, wherein said protein is D. melanogaster, D. virilis, mouse or human.
7. A recombinant isolated Ksr protein produced by a cell comprising a nucleic acid encoding a kinase suppressor of ras (Ksr) according to claim 6 operably linked to a transcription regulatory region not naturally lined to a Ksr-encoding gene.
- Yao et al, 1995, Nature 378, 307-310; Phosphorylation of Raf by ceramide-activated protein kinase. Dent et al., 1995, Science 268, 1902-1906; Reversal of Raf-1 Activation by Purified and Membrane-associated Protein Phosphatases. Dent and Sturgill, 1994, PNAS USA 91, 9544-9548; Activation of (His)6-Raf-1 in vitro by partially purified plasma membranes from v-Ras-transformed and serum-stimulated fibroblasts. Daum et al., 1994, TIBS 19, 474-480; The ins and outs of Raf kinases. McCormick, 1994, TICB 4, 347-350; Raf: the Holy Grail of Ras Biology. Downward et al. (1995). Cell 83: 831-834, Dec. 15, 1995.
Filed: Dec 13, 1995
Date of Patent: Dec 23, 1997
Assignee: Regents of the University of California (Oakland, CA)
Inventors: Gerry Rubin (Berkeley, CA), Marc Therrien (Union City, CA), Henry Chang (Berkeley, CA), Felix Karim (El Cerrito, CA), David Wassarman (San Francisco, CA)
Primary Examiner: Robert A. Wax
Assistant Examiner: Tekchand Saidha
Attorney: Richard Aron Osman
Application Number: 8/571,758
International Classification: C12N 912; C12P 2106; C07K 100; C07H 2104;